基于环介导恒温扩增法快速检测破伤风梭菌

Rapid detection of Clostridium tetani by loop-mediated isothermal amplification assay

目的利用环介导恒温扩增技术,设计快速检测破伤风梭菌的方法,以便对破伤风梭菌感染进行快速检测。方法 (1)设计针对破伤风梭菌的环介导恒温扩增检测方法。(2)对破伤风梭菌恒温扩增法进行特异性试验。(3)对破伤风梭菌恒温扩增法进行灵敏度试验。结果 (1)设计出针对破伤风梭菌的恒温扩增检测方法。(2)本恒温扩增检测方法只扩增破伤风梭菌DNA,对其他菌不扩增,显示出良好的特异性。(3)本恒温扩增检测方法的最低检测限为1×101 CFU/25μL,灵敏度高。结论本试验设计的破伤风梭菌恒温扩增检测方法具有较高的灵敏度和特异性,简便、快速,适用于破伤风梭菌现场检验。

Objective To design a method,using loop-mediated isothermal amplification（LAMP）,for clinical point-of-care testing（POCT） of Clostridium tetani infection.Methods Detection method,based on LAMP,was designed and the specificity and sensitivity of the constructed method were analyzed.Results The LAMP detection system of Clostridium tetani was designed successfully,which was with high specificity and sensitivity（with the lowest detection limit of 1×101 CFU/25 μL） for the detection of Clostridium tetani.Conclusion The LAMP assay of Clostridium tetani,constructed in this research,showed high specificity and sensitivity,with the advantages of convenience and fast detection,and could be used for the POCT of the Clostridium tetani infection.